Berglund:Buffers

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Chemicals

Deionization of Formamide
(From short protocols pg. A1-17) Mix 50ml of formamide and 2.5 grams of mixed-bed ion-exchange resin and stir 45 min. at room temperature. Filter through whatman filter paper, dispense into 30 ml aliquots which are stored at -20° C. Handle formamide with care, it is a teratogen. Deionized formamide lasts for 1 yr. if stored at -20° C. Mixed-bed ion exchange resin will work to remove ions from water, also.

DEPC water
1ml of DEPC in 1L of Millique water. Shake vigorously, let stand in hood overnight, autoclave for 45min. with loosened caps in shallow pan. Allow to cool and screw down caps.

Akta

S column Buffer A (low salt)
25ml 1M HEPES pH 7.5, 20ml 5M NaCl, 2ml 0.5M EDTA pH 8, 0.5ml 2M DTT. 952.5ml nanopure water. Mix all but DTT together. Filter, then add DTT. Mix well. Store in fridge

S column Buffer B (High salt)
12.5ml 1M HEPES pH 7.5, 120ml 5M NaCl, 1ml 0.5M EDTA pH 8, 0.25ml 2M DTT. 366.25ml nanopure water. Mix all but DTT together. Filter, then add DTT. Mix well. Store in fridge

Sizing Buffer
10mM Mes pH 7.5, 300mM NaCl, 1mM EDTA, 1mM DTT

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